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Cardiosphere-Derived Cells from Not Dilated and Dilated Human Myocardium Exhibit Enhanced Metabolic Potential
Daiva Bironaite1, Rokas Mikšiūnas1
1Department of Regenerative Medicine, State Research Institute Centre for Innovative Medicine, LT08406 Vilnius, Lithuania.
Insights
Cardiac sphere-derived cells (SDCs) show improved mitochondrial function and cardiac progenitor gene expression compared to human mesenchymal stem cells (hmMSCs). This suggests SDCs hold promise for treating dilated cardiomyopathy (DCM).
Area of Science:
- Cardiovascular Biology
- Stem Cell Biology
- Metabolic Engineering
Background:
- Dilated cardiomyopathy (DCM) is a leading cause of heart failure and necessitates cardiac transplantation.
- Human myocardium-derived mesenchymal stem/stromal cells (hmMSCs) are being explored for regenerative therapies.
- Understanding the metabolic and progenitor potential of stem cells derived from healthy versus pathological myocardium is crucial.
Purpose of the Study:
- To investigate and compare the metabolic potential of hmMSCs and cardiac sphere-derived cells (SDCs).
- To analyze progenitor and cardiac commitment profiles of SDCs derived from healthy and dilated myocardium.
- To evaluate the suitability of SDCs for potential therapeutic applications in DCM.
Main Methods:
- Isolation and expansion of hmMSCs from myocardial tissues.
- Cultivation of hmMSCs to generate SDCs.
- Analysis of cell morphology, proliferation, mitochondrial activity (Seahorse assays), intracellular calcium levels, and gene expression (qPCR).
Main Results:
- SDCs (both healthy and pathological) exhibited significantly enhanced mitochondrial function compared to hmMSCs.
- SDCs demonstrated increased maximal respiration, ATP production, and coupling efficiency.
- SDCs showed reduced steady-state intracellular calcium levels and upregulated cardiac progenitor/lineage-commitment genes.
Conclusions:
- SDCs possess a superior metabolic, progenitor, and cardiac commitment profile compared to conventional hmMSCs.
- SDCs derived from both healthy and dilated myocardium show promising characteristics for regenerative medicine.
- Further enhancement of metabolic potential in hmMSCs and SDCs from dilated myocardium may be achievable via 3D culture.
Abstract:
Dilated cardiomyopathy (DCM) is a major contributor to heart failure and cardiac transplantation. This study investigated the metabolic potential of human myocardium-derived mesenchymal stem/stromal cells (hmMSCs) and subsequently cardiac sphere-derived cells (SDCs) obtained from healthy (non-dilated) and pathological (dilated) myocardial tissues. hmMSCs were isolated using the explant outgrowth method and expanded in monolayer culture. Small round cells loosely attached on hmMSCs were harvested and cultivated as cardiac spheroids for 1-3 days, subsequently obtaining SDCs. The cell morphology, proliferation rate, mitochondrial activity, and intracellular calcium levels were analyzed using flow cytometry, Seahorse metabolic assays, and spectrophotometry, while expression of cell progenitor and cardiac commitment genes were analyzed by quantitative PCR. Both healthy and pathological SDCs demonstrated significantly enhanced mitochondrial function-reflected by increased maximal respiration, ATP production, and coupling efficiency-along with reduced steady-state intracellular calcium levels compared with hmMSCs. SDCs from both healthy and dilated myocardium showed marked upregulation of several cardiac progenitor and lineage-commitment genes relative to hmMSCs. SDCs derived from both healthy and dilated myocardiums possess a more favorable metabolic, progenitor and cardiac commitment profile than conventional hmMSCs. hmMSCs and SDCs from dilated myocardium retain residual metabolic potential, which may be further enhanced under 3D culture conditions.
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